Genome editing refers to the experimental methods to targeted modify specific loci in the genomic DNA sequence. In recent years, engineered endonucleases, including ZFN, TALEN and CRISPR/Cas, have been developed as a new-generation genome editing technique, and greatly improved the feasibility of gene function analyses, gene therapy, etc. Here, we briefly summarize the basic principle, developmental process and applications of this technology.
Clustered Regularly Interspaced Short Palindromic Repeats ; Endonucleases ; genetics ; Genetic Engineering ; methods ; Genetic Therapy ; Genome ; Genomics

ObjectiveTo observe the survival and pathway of regenerative axons of spinal motoneurons following root injury of brachial plexus (BPI) and to investigate the correlation between the pathway of regenerative axons and the recovery of motor function. Methods48 adult male SD rats were randomly divided into four groups: crush group, cut group, avulsion group and the control group. 2 months later, the functional recovery was evaluated with Grooming test. Fluorogold labeled regenerative motoneurons retrogradely when injected into the motor or sensory branch of musculocutaneous nerve. 3 d later, the C5-8 spinal cord were harvested and the regenerative motoneurons were counted. The survival neurons were counted with neutral red staining. ResultsThe number of survival neurons in the cut group (1005±89) were more than that of the avulsion group （881±76）, but less than the crush group (1126±25). The control group was the most (1900±39）. The regenerative motoneurons reinnervated motor branch were much more than that of the sensory branch in all groups. For the Grooming test, the grade in the crush group (3.42) was much higher than that of the avulsion (1.75) and cut (1.75) group. ConclusionRegenerative axons of spinal motoneurons reinnervate the motor branch after BPI. The number of survival neurons is different after different injuries, which associated with the motor functional recovery.

Chronic Disease ; Humans ; Nasal Polyps ; classification ; pathology ; Sinusitis ; classification ; pathology

AIM: To investigate the therapeutic effect of a new warm moist air device assisted with eyelid massage in the treatment of office dry eye (ODE).METHODS: Sixty ODE patients were recruited,and these patients were randomly divided into two groups.One group received artificial tear treatment,and the other group received a new eyelid warm moist air device combined with eyelid massage treatment.Subjects completed the ocular surface disease index (OSDI),which was followed by fluorescein tear breakup time (BUT),corneal fluorescein staining (CFS),Schirmer I test (SⅠt) and meibomian gland secretion function evaluation at baseline,and 2 and 4wk after treatment.RESULTS: In these 60 patients,48 patients(23 in artificial tear treated group and 25 in the new warm moist air device assisted with eyelid massage treated group)were positive for meibomian gland dysfunction (MGD).Compared to artificial tear treatment,the new warm moist air device assisted with eyelid massage treatment showed favorable therapeutic effect on OSDI (FGroup=41.63,P<0.01),BUT (FGroup=60.47,P<0.01) and the grade of meibum quality (FGroup=12.12,P<0.01) during 4wk treatment in this study.No significant difference was found in SIT (FGroup=0.17,P>0.05),CFS (FGroup=1.79,P>0.05) and meibomian gland expressibility (FGroup=0.68,P>0.05) between the two groups during 4wk treatment.CONCLUSION: This new eyelid warm moist air device assisted with eyelid massage has favorable therapeutic effect over artificial tear in ODE treatment.

Objective To analyze the allergens detection in asthmatic infants and young children. Methods The specific IgE test and allergens screening test were conducted in 243 infants and young children with whooping diseases including asthmatic bronchitis, asthmatic pneumonia and asthma by skin prick test and Pharmacia UniCAP test. Results 1. Food allergen was the allergen detection most frequently in infants(P = 0.03). The proportion of aeroallergens increased significantly with age (positive rate:67. 2%). Milk and egg white was the main food allergen detection in infants and young children,and the main aeroallergen detection was house dust mite. 2. The allergen - specific IgE test using UniCAP test system was more accurate than skin prick test. Conclusions The infants are more susceptible to food hypersensitivity; the increasing opportunity of aeroallergen exposure may lead to a higher prevalence of aeroallergens in young children. UniCAP system is more accurate than other methods.

Objective To observe the preventive effect of immune tolerance induced by immature myeloid dendritic cell(iMDC) loading P2 58-73 peptide(P2 58-73aa-iMDC) in experimental autoimmune neuritis(EAN),and the effect on the expression of IFN-? and IL-33 mRNA.Methods(1)The iMDC were cultured in vitro for loading P2 58-73aa.(2) 21 adult female Lewis rats were randomly divided into the EAN(A)group,iMDC(B)group and P2 58-73aa-iMDC(C)group,and subcutaneously injected PBS,iMDC and P2 58-73aa-iMDC respectively.After 7 d,all the rats were immunized with P2 53-78aa and complete freunds adjuvant(CFA) to inducing EAN.Then,the situation of onset were observed and clinical score were evaluated till 16 d(the crest-time of onset) after immunization.(3) The lymphocyte proliferative response were assayed by 3H-TdR incorporation.The expression of IL-33,IFN-? mRNA in sciatic nerves,spleen and lymph node were detected by RT-PCR.Results(1)In the crest-time of onset,the clinical scores in groups A,B and C were(7.4?1.9),(5.2?1.6) and(3.4?0.9) respectively.There were significant differences between each two groups(allP

AlM: To investigate the effect of exogenous peroxisome-proliferator-activated receptor-γcoactivator-1α ( PGC-1α) on vascular endothelial growth factor ( VEGF) expression in human retinal vascular endothelial cells ( HRVEC) .METHODS:Recombinant PGC-1α protein was added to HRVEC, and no recombinant PGC-1α protein was added to HRVEC as control group. After 24h of incubation, two groups of cells were then placed into a normoxic ( 20%O2 ) or hypoxic ( 1% O2 ) environment for another 16h. The expression of VEGF mRNA and protein were detected by real - time PCR, ELlSA and immunofluorescence cytochemistry.RESULTS: VEGF mRNA and protein levels in the cells were significantly increased by recombinant PGC - 1αprotein both under normoxia and hypoxia conditions as compared with control groups (P<0. 01).CONCLUSlON: PGC-1α can upregulate the expression of VEGF in HRVEC under normoxia and hypoxia conditions.

Objective To investigate the effect of matrix metalloproteinase(MMP-2) in the attack of exprimental autoimmune myositis(EAM) rats, and the effects of methylprednisolone on expressions of gene mRNA and protein of MMP-2. Methods The expression levels of MMP-2 in the peripheral blood, spleen lymphocytes and muscles were detected by using RT-PCR and immunohistochemistry techniques and was compared among EAM group,EAM with methylprednisolone treatment group (EAMM) and control group. Results ⑴ The degree of episode in EAMM group was lower than that of EAM group , and the infiltration of the inflammatory cells and the necrosis of the muscles were also mild in EAMM group. ⑵There was significant upregulation of the expression of the MMP-2 gene mRNA in the lymphocytes of peripheral bood and spleen of EAM rats as compared with that of the control,Elevation of the expression of MMP-2 protein in rats muscles tissues of EAMM group was observed obviously. However, the expression of the mRNA and the protein of MMP-2 was suppressed significantly in EAM group compared with control group. Conclusions The upregulation of the expression of MMP-2 may be correlated with the onset of EAM. Methylprednisolone may relieve the degree of the pathology of EAM by the downregulation of MMP-2 .

To discuss the protective effect of aralosides (AS) on I/R-induced rat myocardial injury. The adult rat ventricular myocyte ischemia model was established through perfusion with sodium lactate perfusate and reperfusion with Ca(2+) -containing Tyrode's solution simulation. The cell contraction and ion concentration synchronization determination system was applied to detect the effect of AS on single I/R cell contraction and Ca2+ transients. According to the findings, AS could increase resting sarcomere length, contraction amplitude, ± dL/dt(max), calcium transient amplitude and speed of post-reperfusion myocardial cells (P < 0.05, P < 0.01), and decrease in time for achieving 90.0% of maximum relaxation, time for achieving peak value, resting calcium ratio, contraction period [Ca2+] i, time for achieving 50.0% of maximum relaxation and attenuation rate of intracellular calcium transient (P < 0.05, P < 0.01). Therefore, it is suggested that AS improved the post-reperfusion cell contraction and injury of calcium homeostasis.
Animals ; Aralia ; chemistry ; Biological Transport ; drug effects ; Calcium ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Muscle Contraction ; drug effects ; Myocardial Ischemia ; drug therapy ; metabolism ; physiopathology ; surgery ; Myocardial Reperfusion ; Myocytes, Cardiac ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Saponins ; administration & dosage

Adenoma, Oxyphilic ; classification ; pathology ; Carcinoma, Renal Cell ; classification ; metabolism ; pathology ; Humans ; Kidney ; pathology ; Kidney Neoplasms ; classification ; genetics ; pathology ; therapy ; Molecular Biology ; methods ; trends